Chicago-Seq of Thielaviopsis punctulata DSM102798
Chicago libraries were prepared as described previously (Putnam et al,2016). Briefly, for each library, ~500ng of high molecular weight genomic DNA (gDNA)with mean fragment length = 100 was reconstituted into chromatin in vitro and fixed withformaldehyde. Fixed chromatin was digested with DpnII, the 5 overhangs filled in withbiotinylated nucleotides, and then free blunt ends were ligated. After ligation, crosslinkswere reversed and the DNA purified from protein. Purified DNA was treated to removebiotin that was not internal to ligated fragments. The DNA was then sheared to ~350 bpmean fragment size and sequencing libraries were generated using NEB Next Ultraenzymes and Illumina-compatible adapters. Biotin-containing fragments were isolatedusing streptavidin beads before PCR enrichment of each library. The libraries weresequenced on an Illumina HiSeq X.